Please let me know sample from which you are working

Hello,

I use the Qaigen RNeasy and it works fine:

http://www.qiagen.com/knowledge-and-support/resource-center/resource-download.aspx?id=14e7cf6e-521a-4cf7-8cbc-bf9f6fa33e24&lang=en

Protocol involves membrane disruption and homogenisation using a Qaizol (i.e. Phenol with guanidine salts) and chloroform. Precipitated RNA is separated by centrifugation and adhered to a silica membrane with 70% ethanol. It's useful to add DEPC to the 70% ethanol to eliminate residual RNases.

After 2 steps of washes and an elution, you are sure to get a fair yield of RNA!

I am 100% agree with Oluwapelumi Adeyemi... RNAeasy kit from Qiagen is working fine... I also isolated RNA from cells from this kit and that works fine.

Thanks for the paper Andrei. It's very relevant. I am also mostly interested in poly-A+ RNAs so this is no problem.

Matthias, you are very right. I sure don't want all RNAs. I My point was that I am not looking for a specific RNA molecule or a specific protein so I can not fish these out. I am interested in long non-coding RNAs that are interacting directly with proteins. But as you say, I guess they all are... So probably there is no such easy procedure to do this. Furthermore, I guess most of these protocols identify RNA-binding proteins, whereas I am interested in protein-binding RNAs...

Hi Jon,

Check out papers cited below and references therein:

Tsai BP, Wang X, Huang L, Waterman ML. Quantitative profiling of in

vivo-assembled RNA-protein complexes using a novel integrated proteomic approach.Mol Cell Proteomics. 2011 Apr;10(4):M110.007385. doi: 10.1074/mcp.M110.007385.

Iioka H, Loiselle D, Haystead TA, Macara IG. Efficient detection of

RNA-protein interactions using tethered RNAs. Nucleic Acids Res. 2011

Apr;39(8):e53. doi: 10.1093/nar/gkq1316. Epub 2011 Feb 7. PubMed PMID: 21300640;

PubMed Central PMCID: PMC3082893

Walker SC, Scott FH, Srisawat C, Engelke DR. RNA affinity tags for the rapid

purification and investigation of RNAs and RNA-protein complexes. Methods Mol

Biol. 2008;488:23-40. doi: 10.1007/978-1-60327-475-3_3. PubMed PMID: 18982282;

PubMed Central PMCID: PMC2807123.

Dear jon

You can look for this article

Hmgb1 can upregulated CD14 monocyte cells via upregulating TLR4